Hormone derivative



' and hydrogen pressure.

Patented Jan. 8, 1946 UNITED STATES PATENT OFFICE 2,392,660 HORMONEDERIVATIVE Gordon A.Grant, Montreal, Quebec, and Carl von Seemann,Westmount, Quebec, Canada No Drawing. Application July 19, 1944. SerialNo. 545,740. In Canada August 20, 1943 INTRODUCTION tached to the 1'7carbon atom and a sulphate group attached to the 3 carbon atom, or saltsthereof.

Onnc'rs It is a principal object of the invention to providewater-soluble hormone products having high oral activity. It is afurther object of the invention to provide hydrogenation products of 17keto steroid hormone sulphates.

With these and other objects in view, the applicants have found thathydrogenated sulphates possessing a 17 hydroxyl group can be preparedfrom 1'7 keto steroidoestrogenic sulphates and that these new productshave high oral oestrogenic activity.

Apreferred procedure for the conversion is catalyticv hydrogenation inaqueous solution under neutral or mildly alkaline conditions in thepresence or a suitable catalyst, preferably platinum oxide, atsubstantially normal temperature Surprisingly, under these conditions,the 3 mono-sulphate group remains attached to the nucleus and the yieldof the hydrogenation products may be practically quantitative. Theproducts can be isolated as water-soluble sodium salts or insolublequinidine salts.

The reaction is characterized specifically by the conversion of sodiumoestrone sulphate CmH-nOsNaS to sodium oestradiol 3 mono-sulphateCrsHaaOeNaS, whichis represented by the following equation.

Claims. (.01. 260-2395) OH H:

H LL...

It should, however, be understood that these formulae are given merelyas temporary assumptions and may later be shown to requiremodifications.

EXAMPLES Example 1 100 mg. sodium oestrone sulphate dissolved in ml. M/5phosphate buffer were hydrogenated at room, temperature in the presenceof 25 mg. platinum oxide catalyst (previously saturated with hydrogen atsubstantially normal pressure) until the equivalent of 2 atoms ofhydrogen had been taken up. After separation of the catalyst, thesolution was evaporated to dryness in vacuo at about C. under nitrogen.The resultin dry residue was exhaustively extracted with absolutemethanol at 40 C. and filtered. The filtrate was evaporated to drynessin vacuo under hydrogen and the above procedure was repeated severaltimes. The final salt-free methanol solutlon was concentrated in vacuounder hydrogen to a small volume and precipitated with live volumes oiether. Sodium oestradiol sulphate separated instantly, was collected ona filter and dried in vacuo. The compound gave the following analysis:

S: 8.43%, oestradiol content 71% (Marrianw Kober test) CmHsaOsSNarequired 8.55% s and 72% oestradiol.

A portion of the hydrogenated compound was precipitated from aqueoussolution by a solution so containing 85 mg. quinidine sulphate in 20 ml.water. Quinidine oestradiol sulphate separated immediately and afterstanding for a. time at 4 C.was collectedonaflltelgwashed with ice-coldwater and drledin vacuo. The yield was practically quantitative. Thesalt was found to be remarkably stable. Ana-lads gave thefollowingflguresz S: 4.49%, NGDumas 4.32%: MM requires:

it 4.14%, S. 4.73%. of the hydrogenated compmmd yielded oestradiol.

thmthenm stancgorwhen orallytothe adults-at. Oflaer immanent-tamarind!It will be that. wlflaout departing fromthewiritodtheinvmflmqtheamdthochimsvarlousmybcmdemihe We claim! ltbs'anewcompounisodiumocstradlolummoutradiols 3.Asanew 4.prooessofproducingoshogmicwhichcumtimsubaufllmflt n w m 2jectingancmtainlngal'l 174'0.'lhesodimnoesh'adiolsmono-sulphateaketostemid3 mmo-sulphatetothenotional catalyticallyactivatedinanaoueouswherebytheadphategroupranainsat dcgreeofoesir lm n wfl yt fl n mtachedtothenucleusandfliel'lketogroupisrcducedtoahydroxyl group.

fled oestradlol.

Example 2 200 mg. sodium oestrone sulphate in 40 ml. M15 bull'er weremind in the presence of 50 mg. plaflnum oxidecatalystprevlouslysaturatedwithhydrogenataubstantially normal hydrogenpressure, until the theoretical amount of hydrogen had been taken up.After separation from the catalyst, the solutionwas evaporatedtodrynessinvacuoatabout 40 C. under nitrogen. The dry residue wasexhaustively extracted with absolute methanol, centrifuged, thesupernatants decanted, the insoluble precipitates washed'on thecentrifuge with fresh absolute methanol, and the clear washings unitedto the supernatant obtained above. The combined methanollc extracts wereconcentrated in vacuo under nitrogen to a small volume, and precipitatedwith live volumes of ether. The precipitate was centrifuged It wasthenre-extracted as above with fresh methanol and the above procedure ofprecipitation repeated several times imtil the precipitate of sodiumoestradiol sulphate was completely free from buffer salts. This final opmdudn oesum genichoa'mmesulphatlswhlchsubjectinganoestrogenic,a17

precipitate was dmolved in 25 m1. absolute methsun] and evaporatedinvacuoundernitrogento a volume 01' a few ml. and precipitated with livevolumes of ether. A practically quantitative yield of sodium oestradiolsulphate was thus obtained.

A portlon of the above methanolic solution was evaporated in dryness invacuo under nitrogen. Upon acid hydrolysis in aqueous solution amactically quantitative yield of oestradiol was obtained, M. P. 164-165C. Upon tion, the material gave no depresion with an authentlc sample ofoestradiol (M. P. 1'15 0.) when a mixed melting point was determined.

Emmple 3 A sample of a suitably purified preparation of the oestrogenicsulphates isolated from equine urine was hydrogenated substantially asin the keto steroid 3 m to the action of catalytically activatedhydrogen in a. neutral or mildly alkaline solutlommsdea' subnormalconditionsof temperahn'e and dhvdros pressurew f ythegrouremainsattachedtothenucleusandthel'lketo groupisredueedtoahydroxylgroup.

A pr ss of producing hormone sulphates, which comprism, subjectingsodiumoesiu'onesulphaietotheacfionofcatalyflcaliy activated hydrogen inan aqueous solution, whereby the sulphate group remains to thenucieusand the 17 keto groupisreducedtoa ydro vlgroup.

7. A process of producing hydrogenated hormone sulphates, whichcomprises, subjecting sodium oestrone sulphatetothe action ofcatalyticallyaetivatedhydrogeninaneutraltomildiy alkaline aqueoussolution. under substantially normaloonditionsoftemperahireandofhydrogen pressure, whereby the sulphategroup remalnsattachedtothenucleusandthe l'iketogroupisreducedtoahydroxylgroup.

a. I'll 1 :1:

gmic sulphates, comprising, subjecting a maralaonofal'lketostemidllmono-sulphateoccun-inginequineurinetotheactimiofcatalyflcallyacia'vatedhydrogenmaneutraltomiidlyalkaline aqueous solution under slbstantially normal conditions ortemperature and ofhydrogen,wherebythe17ketogroupisconvertedtoarlhydroxylgrounandthesulphategroupremainsattachedtothenucleus.

9..A asclaimedinclaimiwhereinthe catabstisplafinumoxide.

10.Aprocessofproducingoestrogenic hormone sulphates, which subjecting awater-soluble substance containing a 17 keto steroid 3 mono-sulphate tothe action of catalytlcally activated hydrogen in an aqueous solutionwhereby the sulphate group remains attached to the nucleus and the 1'1keto groupie reduced to a hydroxyl group, and converting the resultingproduct into a salt.

4 GORDON A. GRANT. CARL VON SEEMANN.

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